Screening of Interacting Proteins with Protein Elicitor Hrip1 and Its Prokaryotic Expression

doi:10.13560/j.cnki.biotech.bull.1985.20170001

Abstract

Abstract: The protein elicitor Hrip1 is a novel hypersensitive response-inducing protein secreted by necrotrophic fungus,Alternaria tenuissima,and it induces defense response and enhances broad-spectrum disease resistance in tobacco,Arabidopsis and rice plants. In order to investigate the molecular mechanism of Hrip1 inducing plant disease-resistance,we screened its interaction partners from a rice cDNA library using Hrip1 as bait protein and via yeast two hybrid system. After retransformation assay and x-a-gal staining,Hirp1-interacting protein CSN5 was obtained. Then,the recombinant pGEX-6P-2-CSN5 plasmid was constructed and transformed into the chemical competent cells of Escherichia coli BL21. The recombinant fusion protein in a single band was purified with a GST affinity chromatography column,and identified by Western blot. The results provided a base for further identification of the protein interacted with Hrip1 and its functions.

Key words: protein elicitor, interacting protein, yeast two hybrid system, prokaryotic expression

LI Shu-peng, YANG Xiu-fen, YUAN Jing-jing, QIU De-wen. Screening of Interacting Proteins with Protein Elicitor Hrip1 and Its Prokaryotic Expression[J]. Biotechnology Bulletin, 2017, 33(6): 182-189.

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